LOCUS       sequence-229539-        6811 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mutational dissection of the G rich region of the c-kit promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6811)
  AUTHORS   Ducani C, Bernardinelli G, Hogberg B, Keppler BK, Terenzi A
  TITLE     Interplay of Three G-Quadruplex Units in the KIT Promoter.
  JOURNAL   J Am Chem Soc. 2019 Jul 3;141(26):10205-10213. doi: 
            10.1021/jacs.8b12753. Epub 2019 Jun 20.
  PUBMED    31244182
REFERENCE   2  (bases 1 to 6811)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/jacs.8b12753"; journalName: "J Am Chem Soc"; date: 
            "2019-07-3- 3"; volume: "141"; issue: "26"; pages: "10205-10213"
COMMENT     Sequence Label: pC-KIT8
FEATURES             Location/Qualifiers
     source          1..6811
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1269..1278
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1275..2210
                     /codon_start=1
                     /product="Renilla luciferase"
                     /label=hRluc
                     /note="human codon-optimized"
                     /translation="MASKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAEN
                     AVIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAW
                     FELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDI
                     ALIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPRE
                     IPLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKV
                     KGLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     polyA_signal    2269..2317
                     /note="synthetic polyadenylation signal"
     promoter        2326..3077
                     /label=HSV TK promoter
                     /note="herpes simplex virus thymidine kinase promoter"
     regulatory      3107..3116
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3113..4765
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /db_xref="GI:42718124"
                     /protein_id="AAS38485.1"
                     /translation="MADAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     RBS             4072..4080
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    4809..4930
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4820..4839)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4874..4893
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        5063..5167
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5168..5236,5237..6028)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(5386..5405)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6199..6787
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6688..6707
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggttcctgg ccttttgctg gccttttgct cacatggctc gacagatctg agggatgtct
       61 tattagttta tcagaatgca tagggaaagc tcatctttct tggagtaggt cttccaataa
      121 gtgttcaagc cctgggtctt ccctcactac aataataaac atacctgctc taccctcctc
      181 acaaagctgt tatcgtgagc atgaaaagac gagtcgctgt ggggaagaac tattaacccc
      241 tatacagata ctgttgttgc tttccgttca aggatccacc atgagactgt gggtaaggca
      301 ctttctgggc agtgaaatcg ggcttaagag aatcattgta cttttatttt gctcctggga
      361 ccggcaaggg cagttaagag ccctagccct aaaactgtcg tctgttacag aggcagagaa
      421 atcattactg gggttggcag cgttgggggg aggggggaaa acgtgtatga aaacctgggc
      481 tcccagagca aacttcgcca cgcgccctcc atcgcccctg tctccacaca agtcccggca
      541 ggggcttctt gggcggaggc cacgccgacc aataggaacc cactgtgttc ctacaggtta
      601 cgaagcaggt ggagaaattg agcagaaaca attagcgaaa ccgggctcag cctttaccgc
      661 ggtgccagga gctcctaaca ggcctggagg ggaatgtcgg ggctcaattt cctaacgctc
      721 ccctccccat ccccatgcca cctccacgag cagcggcgtc cagcctcctc ccgcccgaac
      781 gtgctcgagg ggcgggcagt cgacctttat tgtctgggga gcacctggca ggtggcgggc
      841 ccgtgcccta acgtgtgcgt ggtgcccagc ttcacaaagc gagcgggcag cacctccttg
      901 gtccgggaac gcctcagcct ggccgtccac atcccagggg tggaaaggtg gagagagaaa
      961 ggggctccgg agtcaagagc ggggagagag ggcgcgcgcc ctcctcctcc cggcgggcac
     1021 agccccccgg cattaacacg tcgaaagagc aggggccaga cgccgccggg aagaagcgag
     1081 acccaaacaa acgcgaaaaa gaaaagacga gaagaaacgt gaccgacgcg cagaaagaga
     1141 acgctaagag gaggaactgc tgctcgccgc tcgcggctct gggggctcgg ctttgccgcg
     1201 ctcgctgcac ttgggcgaga gctggaacgt ggaccagagc tcggatccca tcgcagctac
     1261 cgcggctagc caccatggct tccaaggtgt acgaccccga gcaacgcaaa cgcatgatca
     1321 ctgggcctca gtggtgggct cgctgcaagc aaatgaacgt gctggactcc ttcatcaact
     1381 actatgattc cgagaagcac gccgagaacg ccgtgatttt tctgcatggt aacgctgcct
     1441 ccagctacct gtggaggcac gtcgtgcctc acatcgagcc cgtggctaga tgcatcatcc
     1501 ctgatctgat cggaatgggt aagtccggca agagcgggaa tggctcatat cgcctcctgg
     1561 atcactacaa gtacctcacc gcttggttcg agctgctgaa ccttccaaag aaaatcatct
     1621 ttgtgggcca cgactggggg gcttgtctgg cctttcacta ctcctacgag caccaagaca
     1681 agatcaaggc catcgtccat gctgagagtg tcgtggacgt gatcgagtcc tgggacgagt
     1741 ggcctgacat cgaggaggat atcgccctga tcaagagcga agagggcgag aaaatggtgc
     1801 ttgagaataa cttcttcgtc gagaccatgc tcccaagcaa gatcatgcgg aaactggagc
     1861 ctgaggagtt cgctgcctac ctggagccat tcaaggagaa gggcgaggtt agacggccta
     1921 ccctctcctg gcctcgcgag atccctctcg ttaagggagg caagcccgac gtcgtccaga
     1981 ttgtccgcaa ctacaacgcc taccttcggg ccagcgacga tctgcctaag atgttcatcg
     2041 agtccgaccc tgggttcttt tccaacgcta ttgtcgaggg agctaagaag ttccctaaca
     2101 ccgagttcgt gaaggtgaag ggcctccact tcagccagga ggacgctcca gatgaaatgg
     2161 gtaagtacat caagagcttc gtggagcgcg tgctgaagaa cgagcagtaa ttctaggcga
     2221 tcgctcgagc ccgggaattc gtttaaacct agagcggccg ctggccgcaa taaaatatct
     2281 ttattttcat tacatctgtg tgttggtttt ttgtgtgagg atctaaatga gtcttcggac
     2341 ctcgcggggg ccgcttaagc ggtggttagg gtttgtctga cgcgggggga gggggaagga
     2401 acgaaacact ctcattcgga ggcggctcgg ggtttggtct tggtggccac gggcacgcag
     2461 aagagcgccg cgatcctctt aagcaccccc ccgccctccg tggaggcggg ggtttggtcg
     2521 gcgggtggta actggcgggc cgctgactcg ggcgggtcgc gcgccccaga gtgtgacctt
     2581 ttcggtctgc tcgcagaccc ccgggcggcg ccgccgcggc ggcgacgggc tcgctgggtc
     2641 ctaggctcca tggggaccgt atacgtggac aggctctgga gcatccgcac gactgcggtg
     2701 atattaccgg agaccttctg cgggacgagc cgggtcacgc ggctgacgcg gagcgtccgt
     2761 tgggcgacaa acaccaggac ggggcacagg tacactatct tgtcacccgg aggcgcgagg
     2821 gactgcagga gcttcaggga gtggcgcagc tgcttcatcc ccgtggcccg ttgctcgcgt
     2881 ttgctggcgg tgtccccgga agaaatatat ttgcatgtct ttagttctat gatgacacaa
     2941 accccgccca gcgtcttgtc attggcgaat tcgaacacgc agatgcagtc ggggcggcgc
     3001 ggtcccaggt ccacttcgca tattaaggtg acgcgtgtgg cctcgaacac cgagcgaccc
     3061 tgcagcgacc cgcttaaaag cttggcattc cggtactgtt ggtaaagcca ccatggccga
     3121 tgctaagaac attaagaagg gccctgctcc cttctaccct ctggaggatg gcaccgctgg
     3181 cgagcagctg cacaaggcca tgaagaggta tgccctggtg cctggcacca ttgccttcac
     3241 cgatgcccac attgaggtgg acatcaccta tgccgagtac ttcgagatgt ctgtgcgcct
     3301 ggccgaggcc atgaagaggt acggcctgaa caccaaccac cgcatcgtgg tgtgctctga
     3361 gaactctctg cagttcttca tgccagtgct gggcgccctg ttcatcggag tggccgtggc
     3421 ccctgctaac gacatttaca acgagcgcga gctgctgaac agcatgggca tttctcagcc
     3481 taccgtggtg ttcgtgtcta agaagggcct gcagaagatc ctgaacgtgc agaagaagct
     3541 gcctatcatc cagaagatca tcatcatgga ctctaagacc gactaccagg gcttccagag
     3601 catgtacaca ttcgtgacat ctcatctgcc tcctggcttc aacgagtacg acttcgtgcc
     3661 agagtctttc gacagggaca aaaccattgc cctgatcatg aacagctctg ggtctaccgg
     3721 cctgcctaag ggcgtggccc tgcctcatcg caccgcctgt gtgcgcttct ctcacgcccg
     3781 cgaccctatt ttcggcaacc agatcatccc cgacaccgct attctgagcg tggtgccatt
     3841 ccaccacggc ttcggcatgt tcaccaccct gggctacctg atttgcggct ttcgggtggt
     3901 gctgatgtac cgcttcgagg aggagctgtt cctgcgcagc ctgcaagact acaaaattca
     3961 gtctgccctg ctggtgccaa ccctgttcag cttcttcgct aagagcaccc tgatcgacaa
     4021 gtacgacctg tctaacctgc acgagattgc ctctggcggc gccccactgt ctaaggaggt
     4081 gggcgaagcc gtggccaagc gctttcatct gccaggcatc cgccagggct acggcctgac
     4141 cgagacaacc agcgccattc tgattacccc agagggcgac gacaagcctg gcgccgtggg
     4201 caaggtggtg ccattcttcg aggccaaggt ggtggacctg gacaccggca agaccctggg
     4261 agtgaaccag cgcggcgagc tgtgtgtgcg cggccctatg attatgtccg gctacgtgaa
     4321 taaccctgag gccacaaacg ccctgatcga caaggacggc tggctgcact ctggcgacat
     4381 tgcctactgg gacgaggacg agcacttctt catcgtggac cgcctgaagt ctctgatcaa
     4441 gtacaagggc taccaggtgg ccccagccga gctggagtct atcctgctgc agcaccctaa
     4501 cattttcgac gccggagtgg ccggcctgcc cgacgacgat gccggcgagc tgcctgccgc
     4561 cgtcgtcgtg ctggaacacg gcaagaccat gaccgagaag gagatcgtgg actatgtggc
     4621 cagccaggtg acaaccgcca agaagctgcg cggcggagtg gtgttcgtgg acgaggtgcc
     4681 caagggcctg accggcaagc tggacgcccg caagatccgc gagatcctga tcaaggctaa
     4741 gaaaggcggc aagatcgccg tgtaataatt ctagagtcgg ggcggccggc cgcttcgagc
     4801 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     4861 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     4921 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     4981 ggaggttttt taaagcaagt aaaacctcta caaatgtggt aaaatcgata aggatccagg
     5041 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     5101 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     5161 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     5221 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     5281 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     5341 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     5401 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     5461 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     5521 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     5581 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     5641 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     5701 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     5761 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     5821 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     5881 tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     5941 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     6001 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     6061 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6121 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6181 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6241 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6301 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6361 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6421 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6481 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6541 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6601 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6661 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6721 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6781 atggaaaaac gccagcaacg cggccttttt a
//
